Korenkova, V., Slyskova, J., Novosadova, V., Pizzamiglio, S., Langerova, L., Bjorkman, J., Vycital, O., Liska, V., Levy, M., Veskrna, K., Vodicka, P., Vodickova, L., Kubista, M., Verderio, P. The focus on sample quality: Influence of colon tissue collection on reliability of qPCR data. Scientific Reports, 6: 29023, 2016. doi: 10.1038/srep29023.
Successful molecular analyses of human solid tissues require intact biological material with well-preserved nucleic acids, proteins, and other cell structures. Pre-analytical handling, comprising of the collection of material at the operating theatre, is among the first critical steps that influence sample quality. The aim of this study was to compare the experimental outcomes obtained from samples collected and stored by the conventional means of snap freezing and by PAXgene Tissue System (Qiagen). These approaches were evaluated by measuring rRNA and mRNA integrity of the samples (RNA Quality Indicator and Differential Amplification Method) and by gene expression profiling. The collection procedures of the biological material were implemented in two hospitals during colon cancer surgery in order to identify the impact of the collection method on the experimental outcome. Our study shows that the pre-analytical sample handling has a significant effect on the quality of RNA and on the variability of qPCR data. PAXgene collection mode proved to be more easily implemented in the operating room and moreover the quality of RNA obtained from human colon tissues by this method is superior to the one obtained by snap freezing.
Laboratory of Gene Expression
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